Isolation of variants of lymphocytopathic retroviruses from the peripheral blood and cerebrospinal fluid of patients with ARC or AIDS

Abstract

LAV/HTLV-III/AAV viruses were isolated from 20 German patients with ARC/AIDS in order to investigate strain variation. Virus was isolated from the peripheral blood and/or cerebrospinal fluid (CSF) in umbilical cord peripheral blood lymphocyte (PBL) cultures. Isolates were identified by their cytopathic effect (CPE), by reverse transcriptase assays on cell-free infected culture supernatant fluid (SNF), and one or more of the following: immunofluorescence assays on infected cells for viral antigen using HTLV-III reference sera, Western blot analysis of cell-free infected culture SNF, electron microscopy of infected cells, and Southern blot restriction analysis and specific HTLV-III probing of DNA extracted from infected cultured PBL. The isolates could be classified into three groups according to differences in growth rate and cytopathic effect: Most showed what was regarded as the typical CPE, while some either grew rapidly and induced a striking CPE and others grew slowly with minimal CPE. In one patient, virus producing typical CPE was isolated from the peripheral blood while the isolate from his filtered cell-free CSF produced atypical slow CPE, suggesting that antigenic variation may occur with persistent infection or that superinfection may occur. Southern blot DNA restriction analysis of the DNA of three selected isolates showed that two of the isolates were similar but that the restriction pattern of all three differed from patterns previously published. Our results supplement the accumulating evidence of genetic variation among LAV/HTLV-III strains. The extent of this variation needs to be evaluated for any effect on the sensitivity of diagnostic tests, on the strategy of vaccine development, on tissue tropism by altering the viral surface receptor-binding sites, and possibly on the development of specific chemotherapy.

Molecular cloning of two west African human immunodeficiency virus type 2 isolates that replicate well in macrophages: a Gambian isolate, from a patient with neurologic acquired immunodeficiency syndrome, and a highly divergent Ghanian isolate.

“…Primary isolation and cultivation of HIV-2 from patients' peripheral blood lymphocytes (PBLs) cocultured with human umbilical cord blood lymphocytes (CBLs) were done as described (2). For the establishment of permanent virus-producing cell lines, HUT-78 or U937 cells were cocultured with infected CBLs.…”

“…They differ in biological properties such as cell tropism or replication efficiency in a given cell system (2)(3)(4)(5)(6), in antigenic determinants leading to different serological responses (7,8), and in their nucleic acid sequence, often reflected in marked restriction-site polymorphism (3,9,10). Multiple variants have been demonstrated to exist within individual patients (2,3,10,11).…”

Improved Coreceptor Usage Prediction and GenotypicMonitoring of R5-to-X4 Transition by Motif Analysis of HumanImmunodeficiency Virus Type 1 env V3 LoopSequences

“…These phenotypes were often found to be associated with differences in growth properties and cytopathicity on peripheral blood mononuclear cells (PBMC) (1,14,46) and in cellular host range (3,48). Ultimately, the difference between the NSI and SI phenotypes was shown to be due largely to the differential use of chemokine receptors as coreceptors for viral entry: NSI viruses predominantly use CCR5, while SI viruses can use CCR5 and CXCR4 or CXCR4 exclusively (2,29,31,52,54).…”

Simian immunodeficiency viruses (SIVs) from eastern and southern Africa: detection of a SIVagm variant from a chacma baboon.

“…After 2 to 3 days, 2 % Lymphocult T-HP (Biotest) was added and the PBMCs were then cocultivated (1 : 1) with CEM-SS cells (Nara et al, 1987 ;Nara & Fischinger, 1988). Cultures were monitored for cytopathic effect and a lentivirus infection was confirmed by monitoring the culture supernatants for reverse transcriptase (RT) activity as previously described (Rubsamen-Waigman et al, 1986). Total cellular DNA was extracted for PCR from 10) RT-positive cultured lymphocytes from two specimens (Bab590a and Bab590b) by the standard phenol-chloroform method (Sambrook et al, 1989).…”